Methods of sensitizing drug resistant microorganisms using methylsulfonylmethane (MSM)

ABSTRACT

Embodiments of the invention relate generally to formulations comprising methylsulfonylmethane (MSM) and one or more therapeutic agents, and uses of such formulations to treat, inhibit and/or sensitize infectious diseases. In several embodiments, such formulations are effective to treat, inhibit and/or sensitize drug-resistant bacterial microorganisms, for example, methicillin-resistant  Staphylococcus aureus  (MRSA).

RELATED APPLICATIONS

This is a continuation of PCT/US2010/054871, filed Oct. 29, 2010, whichwas published in English under PCT Article 21(2), and claims the benefitof U.S. Provisional Application Nos. 61/256,935, filed Oct. 30, 2009,and 61/319,203, filed Mar. 30, 2010. The disclosure of each of thesepatent documents is expressly incorporated by reference herein.

BACKGROUND

1. Field of the Invention

Embodiments of the invention relate generally to formulations comprisingdimethyl sulfoxide (DMSO) alone or in combination withmethylsulfonylmethane (MSM) to treat infectious diseases. Certainembodiments relate to sensitizing drug-resistant microbes to drugs.Several formulations disclosed herein are useful for treatingdrug-resistant tuberculosis.

2. Description of the Related Art

Infectious diseases are diseases caused by pathogenic microbial agents,including viruses, bacteria, fungi, parasites, and prions, among others.The pathogenic agents may be primary or opportunistic pathogens. Primarypathogens cause infection as a direct result of their virulence, whileopportunistic pathogens typically require a compromised host defensesystem to produce an infection. Examples of common infectious diseasesinclude HIV/AIDS, measles, tetanus, tuberculosis, malaria, upper andlower respiratory infections, and hepatitis. While modern medicine hasreduced the prevalence of many infectious diseases, particularly indeveloped countries, they still account for a large degree of morbidityand mortality.

Tuberculosis, malaria, HIV/AIDS, and diarrhoeal diseases are the leadingkillers among the infectious diseases. Tuberculosis (tubercle bacillus)is caused by mycobacteria, primarily Mycobacterium tuberculosis.Tuberculosis primarily infiltrates the lungs (pulmonary tuberculosis),but has also been documented as affecting the central nervous system,the lymphatic system, and the circulatory system, among others. Othermycobacteria may also cause tuberculosis, for example, Mycobacteriumbovis, Mycobacterium africanum, Mycobacterium canetti, and Mycobacteriummicroti. However, these species are less common in humans.

Despite certain improvements in medical treatment for infectiousdiseases (antibiotics and vaccines), there remain many obstacles toreducing the mortality caused by these diseases. A primary issue is theemergence and spread of drug resistant pathogens. Tuberculosis and otherinfectious diseases have shown resistance to first-line drugs. In someinstances, resistance has progressed to require a change to moreexpensive second or third-line agents. Evolution of pathogenicmicroorganisms is expected due to the selective pressure of drugtherapy. However, resistance is exacerbated by several factors,including abuse, underuse or misuse of antimicrobials, poor patientcompliance, and poor quality of available drugs.

Dimethyl sulfoxide (DMSO; (CH₃)₂(SO)) is a polar, aprotic solvent widelyused as a solvent. It is frequently used in various chemical andbiological reactions and as a cryoprotectant for cell storage. Thestrong unpleasant odor of DMSO (or metabolites), among other sideeffects, has adversely impacted the use of DMSO in medical applications.

Methylsulfonylmethane (MSM; (CH3)₂SO₂)), also known as dimethyl sulfone,is an organosulfur compound that is a metabolite of DMSO and certainsulfur-containing amino acids. MSM has been marketed primarily as adietary supplement.

SUMMARY

There exists a need for an effective and easily administered therapyagainst infectious diseases, particularly those that are developing orhave developed some degree of antimicrobial resistance.

Tuberculosis classically presents as a chronic cough with blood-tingedsputum, fever, night sweats, and weight loss. Tuberculosis may alsoinfect other organ systems, leading to a diverse set of symptoms.Diagnosis of tuberculosis is typically achieved through radiology,tuberculin skin test, blood tests, as well as microscopic examinationand microbiological culture of bodily fluids. Current tuberculosistreatment requires long courses of multiple antibiotics. One course oftreatment for tuberculosis is isoniazid, rifampicin, pyraziamide, andethambutol for about two months, followed by only isoniazid andrifampicin for a further four months. The patient is considered cured atsix months (although there is still a relapse rate of 2 to 3%). Latenttuberculosis therapy involves six to nine months of isoniazid alone.

Drug-resistant tuberculosis includes tuberculosis that is resistant toat least one anti-tuberculosis drug. Multidrug-resistant tuberculosis(MDR tuberculosis) includes tuberculosis that is resistant to more thanone first line anti-tuberculosis drug.

In several embodiments, formulations comprising DMSO alone or DMSO incombination with MSM, and a therapeutic agent are provided to treattuberculosis, including drug-resistant tuberculosis.

In several embodiments, formulations comprising DMSO alone or incombination with MSM are provided as an inhalant to treat drug-resistanttuberculosis. In some embodiments, formulations comprising DMSO alone orin combination with MSM are formulated as solids, while in several otherembodiments, formulations comprising DMSO and MSM are formulated asliquids. In some embodiments, the formulations are consumed orally totreat drug-resistant tuberculosis, while in some other embodiments, theformulations are applied topically. In several embodiments,drug-resistant diseases other than tuberculosis are treated withformulations comprising DMSO alone or in combination with MSM.

In yet other embodiments, non-drug-resistant diseases are treated withformulations comprising DMSO alone or DMSO in combination with MSM. Insuch embodiments, DMSO or a combination of DMSO and MSM is combined witha therapeutic agent to enhance the effects of the therapeutic agent.

In several embodiments, subjects with drug-resistant tuberculosis aretreated with a DMSO formulation, an MSM formulation, or combination ofDMSO and MSM, together with isoniazid, rifampicin, pyrazinamide, andethambutol for a time period (e.g., two weeks-two months), thenisoniazid and rifampicin alone for another time period (e.g., fourweeks-four months). In some embodiments, subjects with drug-resistanttuberculosis are treated with a DMSO formulation, alone or incombination with MSM, together with isoniazid, rifampicin, pyrazinamide,and/or ethambutol. In one embodiment, DMSO alone or in combination withMSM formulations sensitize drug-resistant tuberculosis to antibiotics,and therefore isoniazid, rifampicin, pyrazinamide, and/or ethambutolbecome lethal to sensitized tuberculosis bacteria. In other embodiments,subjects with drug-resistant tuberculosis are treated with a DMSOformulation, alone or in combination with MSM, together with one or moreof the following: isoniazid, rifampicin, pyrazinamide, ethambutol,lamprene, mycobutin, seromycin, streptomycin, myambutol, priftin andrifamate

In several embodiments, a system for treating drug-resistanttuberculosis is provided. In one embodiment, the system comprises atherapeutic agent and DMSO alone, or a combination of DMSO and MSM. Theconcentration of the DMSO and/or MSM ranges from about 15% to about 95%in a total volume of about two to about six milliliters. The therapeuticagent is selected from one or more compounds selected from the groupconsisting of: isoniazid, rifampicin, pyraziamide, and ethambutol. Inone embodiment, the system includes an inhalant device, wherein theinhalant device is constructed from a material adapted for contactingDMSO and/or MSM and wherein the inhalant device is configured foradministering the formulation to a subject. The inhalant device isconfigured to deliver the formulation to directly contact drug-resistanttuberculosis bacteria in the subject's lung tissue. In one embodiment,the DMSO and/or MSM sensitizes the drug-resistant tuberculosis bacteriato at least one of isoniazid, rifampicin, pyraziamide, and ethambutol togenerate sensitized tuberculosis bacteria. The sensitized tuberculosisbacteria are lethally inhibited by exposure to least one of isoniazid,rifampicin, pyraziamide, and ethambutol.

In several embodiments, the invention comprises a method for treatingdrug-resistant tuberculosis. In one embodiment, the method comprisesproviding a formulation comprising a therapeutic agent and DMSO alone,or a combination of DMSO and MSM. In one embodiment, the concentrationof DMSO and/or MSM ranges from about 15% to about 95% in a total volumeof about two to about six milliliters. In one embodiment, thetherapeutic agent is selected from one or more compounds selected fromthe group consisting of: isoniazid, rifampicin, pyraziamide, andethambutol. In some embodiments, the method further includes providingan inhalant device and administering the formulation via the inhalantdevice to a subject having drug-resistant tuberculosis. The inhalantdevice is configured to deliver the formulation to directly contactdrug-resistant tuberculosis bacteria in the subject's lung tissue. Thedrug-resistant tuberculosis bacteria are sensitized (with the DMSO) toat least one of isoniazid, rifampicin, pyraziamide, and ethambutol togenerate sensitized tuberculosis bacteria, and lethally inhibited by atleast one of isoniazid, rifampicin, pyraziamide, and ethambutol. In oneembodiment, a method of treating drug-resistant tuberculosis in thefield is provided. In one embodiment, a test for tuberculosis isperformed within hours of seeing a subject, a test for INH and RRP isgiven within hours, the subject is treated with DMSO alone or incombination with MSM (e.g., intravenously), and then treated with ainhaled formulation of DMSO alone or in combination with MSM, providedwith mask and trained to use an inhalant device in the field. Methods oftreating drug-resistant diseases other than tuberculosis with DMSO aloneor in combination with MSM are also provided according to severalembodiments.

In several embodiments, the invention comprises a formulation forsensitizing drug-resistant microorganisms to a therapeutic agent. In oneembodiment, the formulation comprises a therapeutic agent and DMSO aloneor a combination of DMSO and MSM. The concentration of the DMSO and/orMSM ranges from about 15% to about 95% in a total volume or weight ofthe formulation. The DMSO or combination of DMSO and MSM is configuredfor contacting a drug-resistant microbe, wherein the drug-resistantmicrobe is resistant to the therapeutic agent. After contact with theDMSO or combination of DMSO and MSM the drug-resistant microbes aresensitized to the therapeutic agent and are transformed into sensitizedmicrobes, wherein the sensitized microbes are inhibited by thetherapeutic agent. The drug-resistant microorganism contacted with DMSOor a combination of DMSO and MSM includes, but is not limited to,tuberculosis, malaria, MRSA and streptococcus.

In some embodiments, the inhalant device is an inhaler, nebulizer orventilator. According to several embodiments, the inhalant device isconstructed from a material adapted for contacting DMSO and/or MSMwithout degrading into the subject's airway, trachea, bronchial tubes,or lung tissue, or without rupturing. In one embodiment, the formulationis pre-dosed in the inhalant device.

In some embodiments, the total formulation volume is about 2-8 ml, e.g.,about 3 ml for an inhaler or 7 ml for a nebulizer. According to someembodiments, the formulations described herein also comprise urea. Ureamay be provided in a dose about 1 mg to about 10 grams or higher inseveral embodiments. In one embodiment, formulations disclosed hereinare configured for delivery at least three times daily.

In one embodiment, the formulation comprises a daily dose of: rifampicinin an amount ranging from about 500 mg to about 700 mg, isoniazid in anamount ranging from about 200 mg to about 400 mg, pyrazinamide in anamount ranging from about 2.0 g to about 3.0 g, and ethambutol in anamount ranging from about 1.0 g to about 2.0 g. In another embodiment,the formulation comprises a total daily dose or a daily dose per kg ofbody weight of: rifampicin in an amount ranging from about 1 mg to about100000 mg, isoniazid in an amount ranging from about 2 mg to about 10000mg, pyrazinamide in an amount ranging from about 0.02 g to about 10.0 g,and ethambutol in an amount ranging from about 0.01 g to about 10.0 g.In one embodiment, the formulation comprises a daily dose of about 600mg rifampicin, 300 mg isoniazid, 2.4 g pyrazinamide, and 1.2 gethambutol. Streptomycin or other therapeutic agents are included inseveral embodiments. In one embodiment, streptomycin is provided in atotal daily dose or a daily dose per kg of body weight of about 10 mg toabout 200 g.

A system according to any one of the preceding claims, furthercomprising a pretreatment composition, wherein the pretreatmentcomposition comprises about 50 mg to about 60 mg DMSO and is configuredfor intravenous administration to the subject.

In several embodiments, wherein the inhalant device is configured togenerate particles of the formulation that range in size from about 0.5micron (μm) to about 5 μm.

In one embodiment, the invention comprises a kit for treatingdrug-resistant tuberculosis, comprising: a system according to any oneof the embodiments described herein; and instructions for administeringthe formulation via the inhalant device at least three times daily. Thekit may also include a mask, a battery charger and/or a battery pack.

DETAILED DESCRIPTION

In several embodiments described herein, formulations comprising DMSOalone or in combination with MSM are provided. In several embodiments,such formulations are used to treat tuberculosis, includingdrug-resistant tuberculosis. Despite the increasing prevalence ofdrug-resistant pathogenic microbes, several embodiments of theformulations disclosed herein are unexpectedly effective in treatingdrug-resistant bacteria or other microbes. Other drug-resistantpathogenic microbes are also treated by DMSO alone, or a combination ofDMSO and MSM, together with a therapeutic agent.

Formulations according to several embodiments herein include DMSO alone(e.g., (i) without MSM or any other active ingredient; (ii) without MSMbut with one or more active ingredients; and/or (iii) without MSM andwith or without one or more inactive ingredients). Formulationsaccording to several embodiments herein include DMSO in combination withMSM, wherein the combination may or may not include one or more inactiveor active ingredients.

In several embodiments, the combination of DMSO and MSM allows a lowerconcentration of DMSO and/or MSM to be used. In other embodiment, theuse of DMSO alone or in combination with MSM reduces the minimumefficacious concentration of other constituents of the formulation,thereby also reducing side effects from those constituents. For example,in one embodiment, the addition of DMSO, MSM or DMSO and MSM will permita reduced dosage of antibiotics, or other antimicrobial, to achievecomparable or enhanced therapeutic effects.

In several embodiments, formulations comprising DMSO alone or incombination with MSM sensitize drug-resistant tuberculosis to drugs. Inone embodiment, formulations comprising DMSO alone or in combinationwith MSM partially or fully reverse the drug-resistant nature ofbacterial strains, including tuberculosis.

In one embodiment, DMSO or combination of DMSO and MSM enhances thepermeability of bacteria (or other microbes), thus enhancing theeffectiveness of the antibiotic (or other therapeutic agent). In anotherembodiment, DMSO or combination of DMSO and MSM serves as a carrier thatcarries an antibiotic (or other therapeutic agent) into the cell. In afurther embodiment, DMSO creates permanent or temporary pores on or inthe cell to enhance delivery of drugs. In other embodiments, the use ofMSM in combination with DMSO enhances the effectiveness of DMSO. In someembodiments, MSM reduces the concentration of DMSO required, therebyreducing concomitant side-effects. In further embodiments, MSM incombination with DMSO enhances the permeability of antibiotics and/orserves as a carrier for antibiotics (or other therapeutic agents).

In several embodiments, formulations comprising DMSO or DMSO incombination with MSM and at least one therapeutic agent lower theconcentration of DMSO, MSM, and/or the therapeutic agent needed toeffectively treat one or more types of infection. In one embodiment, aformulation comprising DMSO alone or in combination with MSM sensitizesthe bacteria (whether drug-resistant or not) to antibiotics. Thus, sucha formulation: (i) reduces the dose of antibiotic needed; (ii) reducesthe treatment time; (iii) reduces the number of different antibioticsneeded, and/or (iv) makes an antibiotic effective. Accordingly,undesired side effects associated with antibiotics may be reduced inseveral embodiments, including liver damage, kidney damage, oculardefects, hyperuricemia, thrombocytopenia, leukopenia, and neutropenia.In one embodiment, a DMSO formulation or combination of DMSO with MSMformulation sensitizes drug-resistant tuberculosis to isoniazid,rifampicin, pyrazinamide, and/or ethambutol. In additional embodiments,DMSO alone or in combination with MSM enhances the effects of isoniazid,rifampicin, pyrazinamide, and/or ethambutol on non drug-resistanttuberculosis. Pulmonary and extra-pulmonary tuberculosis are treatedaccording to several embodiments.

Many infections lead to local inflammation (or even inflammation of alarge area of tissue around the site of infection). In some embodiments,DMSO alone or in combination with MSM works synergistically withtherapeutic agents to reduce inflammation to a greater degree than DMSO,MSM or the agent alone.

As used herein, the term “therapeutic agents” shall be given itsordinary meaning and shall include an agent or agents that ameliorate orcure an illness or symptoms of an illness. Therapeutic agents shallinclude, but not be limited to, analgesics, anti-inflammatory agents,antimicrobial agents (such as antiparasitic, antifungal, antiviral, andantibiotic agents), and combinations thereof.

In several embodiments, DMSO and MSM in a single formulation with atherapeutic agent act synergistically to reduce the amount of DMSOneeded to achieve efficacious amounts of therapeutic agent delivery to atarget site of infection. In some embodiments, MSM enhances thepenetrant effect of DMSO, allowing a therapeutic agent to reach a targetarea of infection at an increased concentration (or reduced time frame).Thus, in some such embodiments, the synergy between MSM and DMSO reducethe side effects associated with DMSO administration, which include anunpleasant odor post administration, nausea, diarrhea, and skin/throatirritation, among others.

In still other embodiments, DMSO and MSM in a single formulation with atherapeutic agent act synergistically to reduce the amount oftherapeutic agent needed to effectively treat an infection. For example,many antibiotics have an established minimum inhibitory concentration(MIC) at which they are effective in reducing or killing certainbacteria (e.g., associated with tuberculosis). In some embodiments, aformulation comprising DMSO and MSM and a sub-MIC concentration of anantibiotic is equally or more effective at reducing or killing suchbacteria as compared to the antibiotic alone at MIC levels. In otherembodiments, a formulation comprising DMSO, MSM and an antibiotic aremore effective at reducing or eliminating bacteria associated withtuberculosis from a site of infection as compared to the antibioticalone. In several embodiments, formulations disclosed herein augmenttreatment of multiple drug-resistant tuberculosis. In one embodiment,formulations disclosed herein sensitize mycobacterium tuberculosisstrains in vitro, in macrophages and in vivo to drugs. In oneembodiment, a formulation comprising DMSO and ethambutol sensitizesbacteria to drugs other than ethambutol. In some embodiments,formulations comprising DMSO enhance the sensitivity or susceptibilityto drugs such as sub-MIC concentrations of ethambutol, isoniazid,rifampicin, and streptomycin by about 2-fold to about 100-fold. Unlikeprior reports, concentrations of DMSO greater than 50% are particularlyadvantageous in some embodiments. Enhancement of drug susceptibility ofmulti-drug resistant strains of Mycobacterium tuberculosis by ethambutoland dimethyl sulphoxide. Jagannath et al. Journal of AntimicrobialChemotherapy (1995) 35, 381-390, herein incorporated by reference.

In some embodiments, DMSO alone or in combination with MSM allowantibiotics (or other therapeutic agents) to penetrate lung tissueinfected with tuberculosis. In one embodiment, DMSO alone or incombination with MSM: (i) allow antibiotics to reach deeper levels oftuberculosis infected tissue; (ii) allow direct contact of tuberculosisinfected tissue; (iii) lengthen the exposure time of the antibiotic tothe tuberculosis infected tissue; and/or (iv) decrease the time toachieve a desired antibiotic effect. In one embodiment, DMSO alone or incombination with MSM achieves one or more of these desired effectsthrough use as an inhalant, wherein the inhalant additionally comprisesone or more antibiotics or other therapeutic agents.

In several embodiments, the combined use of MSM reduces or eliminatesthe odor normally associated with DMSO. This is surprisingly beneficialin several embodiments because practitioners have avoided using DMSO inhigh concentrations (or in any amount) because of its unpleasant odor.

In some embodiments, DMSO alone or DMSO in combination with MSMformulations comprise antiparasitic agents that are effective intreating infections caused by parasites, such as nematodes, cestodes,trematodes, protozoa, or amoebae.

In some embodiments, DMSO alone or DMSO in combination with MSMformulations comprise antifungal agents that are effective in treatingfungal infections, such as those caused by ringworm, candidiasis, andCryptococcus (cryptococcal meningitis, for example).

In some embodiments, DMSO alone or DMSO in combination with MSMformulations comprise antiviral agents that are effective in treatingviral infections. In some embodiments, specific classes of antiviralagents are used to treat infections caused by a particular type ofvirus. In some embodiments, agents that target HIV, herpes viruses,hepatitis B or C viruses, and influenza viruses are used.

In several embodiments, DMSO alone or DMSO in combination with MSMformulations comprise antibiotics that are effective in treatingbacterial infections by, for example, inhibiting bacterial growth,metabolism, proliferation, activity and/or function. In someembodiments, bacteriostatic antibiotics are used, while in otherembodiments, bactericidal antibiotics are used. In still otherembodiments, both bacteriostatic and bactericidal antibiotics areincorporated into a single formulation comprising DMSO and/or MSM. Insome embodiments, antibiotics of one or more classes are incorporatedinto a formulation comprising DMSO alone or DMSO in combination withMSM. In certain embodiments, a formulation includes one or more of an:aminoglycoside, ansamycin, carbacephem, carbapenem, cephalosporin(1^(st), 2^(nd), 3^(rd), 4^(th), or 5^(th) generation), glycopeptides,macrolide, monobactam, penicillin, polypeptide, quinolone, sulfonamide,tetracycline, and the like.

In some embodiments, specific diseases are targeted by incorporatingspecific antibiotics into a formulation comprising DMSO alone or DMSO incombination with MSM. For example, macrolides, such as azithromycin orerythromycin are incorporated into formulations used to treatrespiratory or mycoplasmal infections. Likewise, penicillins, such asamoxicillin or oxacillin are incorporated into formulations used totreat a broad range of streptococcal infections.

In still other embodiments, specific disease-causing microorganisms aretargeted by the specific antibiotics incorporated into a formulationcomprising DMSO alone or DMSO in combination with MSM. For example,aminoglycosides, such as neomycin are incorporated into formulationsused to treat Escherichia coli infections. In several embodiments,antibiotics typically used to combat microbial infections are used. Incertain embodiments, antibiotics including, but not limited to,isoniazid, rifampicin, pyraziamide, and ethambutol are incorporated intoformulations comprising one or more of DMSO and MSM, and are used totreat tuberculosis, including drug-resistant tuberculosis.

In several embodiments of the invention, formulations comprising DMSO,MSM and one or more of the following therapeutic agents: rifampicin,isoniazid, pyrazinamide, and ethambutol are provided. In otherembodiments, formulations comprising DMSO and at least one ofrifampicin, isoniazid, pyrazinamide, and ethambutol are provided. Inseveral embodiments, formulations comprising DMSO alone, or incombination with MSM in combination with rifampicin, isoniazid,pyrazinamide, and ethambutol are provided to treat tuberculosis,including drug-resistant tuberculosis.

In some embodiments, rifampicin is provided in a total daily doseranging from about 400 mg to about 800 mg per day. In some embodiments,rifampicin is provided in a total daily dose ranging from about 500 mgto about 700 mg per day, while in still other embodiments, it isprovided in a total daily dose ranging from about 550 to about 650 mgper day, including 560, 570, 580, 590, 600, 610, 620, 630, and 640 mgper day. In some embodiments, higher or lower quantities may be used.

In some embodiments, isoniazid is provided in a total daily dose rangingfrom about 100 mg to about 500 mg per day. In some embodiments,isoniazid is provided in a total daily dose ranging from about 200 mg toabout 400 mg per day, while in still other embodiments, it is providedin a total daily dose ranging from about 250 mg to about 350 mg per day,including 260, 270, 280, 290, 300, 310, 320, 330, and 340 mg per day. Insome embodiments, higher or lower quantities may be used.

In some embodiments, pyrazinamide is provided in a total daily doseranging from about 1.0 to about 4.0 g per day. In some embodiments,pyrazinamide is provided in a total daily dose ranging from about 2.0 toabout 3.0 g per day, while in still other embodiments, it is provided ina total daily dose ranging from about 2.0 to 2.5 g per day, including2.1, 2.2, 2.3, and 2.4 g. In some embodiments, higher or lowerquantities may be used.

In some embodiments, ethambutol is provided in a total daily doseranging from about 0.5 to about 2.5 g per day. In some embodiments,ethambutol is provided in a total daily dose ranging from about 1.0 to2.0 g per day, while in still other embodiments, it is provided in atotal daily dose ranging from about 1.0 to about 1.5 g per day,including 1.1, 1.2, 1.3, and 1.4 g. In some embodiments, higher or lowerquantities may be used.

In some embodiments, DMSO alone or in combination with MSM is used topretreat a patient suffering from an infectious disease, such asdrug-resistant tuberculosis. In some embodiments, the dose of DMSO aloneor in combination with MSM used to pretreat patients ranges from about10% to 50% weight to volume. In some embodiments, the pretreatment DMSOalone or in combination with MSM dose ranges from about 20% to about40%, from about 25% to 35%, including 26, 27, 28, 29, 30, 31, 32, 33,and 34%. In some embodiments, about 50% to about 100% DMSO and/or MSM isused. In several embodiments, pretreatment with DMSO and/or MSM enhancesthe ability of an antibiotic to inhibit tuberculosis associatedbacterial activity and/or sensitizes a drug-resistant strain to a drugthat was previously ineffective.

In some embodiments, a formulation is prepared wherein antimicrobialsare dissolved in DMSO alone or in combination with MSM prior toadministration. This is particularly advantageous in certain embodimentsbecause the antimicrobial and DMSO (and optionally MSM) can beadministered to a subject via inhalation to treat tuberculosis.Inhalants, according to some embodiments, provide direct access of theDMSO alone or in combination with MSM to infected lung tissue tosensitize bacterial cells to the antibiotic to treat tuberculosis.

In one embodiment, an inhalant is provided to target the site ofinfection (e.g., lungs) of several infectious diseases, such astuberculosis. In some such embodiments, the inhalant device comprises anebulizer. In other embodiments, an inhaler is used. In someembodiments, a pressurized metered-dose inhaler is used, and theformulation is inhaled in liquid aerosol form. In other embodiments, drypowder inhalers are used, and the formulation is inhaled in a powderaerosol form. In several embodiments, oral, intravenous, intramuscular,or subcutaneous administration is used in addition to or instead ofinhalant therapy.

The ability to administer antimicrobial agents as an inhalant (e.g., ina powder aerosol form) with DMSO alone or DMSO in combination with MSMis especially advantageous in some embodiments because it allows forincreased shelf-stability and pre-packaged dosages. This is particularlyhelpful for individuals in underdeveloped or developing nations who donot have regular access to healthcare facilities. Entire courses oftreatment can be provided to an affected subject in a single visit to ahealthcare practitioner without the need for a hospital stay or repeatvisits. In several embodiments, formulations disclosed herein aresuitable for self-administration (e.g., through inhalant devices) andare therefore especially appropriate for patients with limited access tohealthcare.

In certain embodiments, the total volume of inhaled DMSO alone or incombination with MSM is about 2-8 ml. In some embodiments, the totalvolume of inhaled DMSO alone or in combination with MSM is about 2 ml toabout 4 ml. In some embodiments, the total volume of inhaled DMSO aloneor in combination with MSM is about 6 ml to about 8 ml. In still otherembodiments, the total volume of inhaled DMSO alone or in combinationwith MSM is about 3 ml to about 7 ml, including 4, 5, and 6 ml. Thus, insome embodiments, the concentration of DMSO administered via inhalationranges from about 65% to about 95%, including 70, 75, 80, 85, 86, 87,88, 89, 90, 91, 92, 93, and 94%, and overlapping ranges thereof.

In several embodiments, MSM is included with the inhaled DMSO andantimicrobial compounds. In certain embodiments, the amount of inhaledMSM used in combination with DMSO ranges from about 0.01% by weight toabout 70% by weight. In other embodiments, the inhaled formulationcontains between about 0.01% and 10% MSM (used in combination with DMSO)by weight. Other embodiments contain between about 10 and 20% MSM, about20-30% MSM, about 30-40% MSM, about 40-50% MSM, about 50-60% MSM, orabout 60-70% MSM including 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, and70% MSM, and overlapping ranges thereof (each used in combination withDMSO). Still other embodiments comprise a formulation containing about 7and 15% MSM, about 15-25% MSM, about 25-35% MSM, about 35-45% MSM, about55-60% MSM, about 60-65% MSM, or about 65-70% MSM (each used incombination with DMSO). Thus, in some embodiments of the inhaledformulation containing MSM, the concentration of DMSO administeredranges from about 50% to about 95%, including 55, 60, 65, 70, 75, 80,85, 86, 87, 88, 89, 90, 91, 92, 93, and 94%, and overlapping rangesthereof.

In several embodiments, the use of MSM reduces the amount of DMSO neededto achieve a comparable effect and/or enhances the efficacy of DMSO byat least 10%, 25%, 50%, 100%, 2-fold, 3-fold, 5-fold, 10-fold, 50-fold,or 100-fold. In other embodiments, the use of MSM reduces the amount ofa therapeutic agent needed to achieve a comparable effect and/orenhances the efficacy of the therapeutic agent by at least 10%, 25%,50%, 100%, 2-fold, 3-fold, 5-fold, 10-fold, 50-fold, or 100-fold. Infurther embodiments, the use of DMSO reduces the amount of a therapeuticagent needed to achieve a comparable effect and/or enhances the efficacyof the therapeutic agent by at least 10%, 25%, 50%, 100%, 2-fold,3-fold, 5-fold, 10-fold, 50-fold, or 100-fold. In still otherembodiments, the use of DMSO and MSM reduces the amount of a therapeuticagent needed to achieve a comparable effect and/or enhances the efficacyof the therapeutic agent by at least 10%, 25%, 50%, 100%, 2-fold,3-fold, 5-fold, 10-fold, 50-fold, or 100-fold as compared to DMSO or MSMalone and/or the therapeutic agent alone.

In several embodiments, a pretreatment formulation comprising DMSO,alone or in combination with MSM, is administered to a subjectintravenously, intramuscularly, topically or orally to enhance theeffects of an inhalant therapy comprising DMSO alone or in combinationwith MSM with therapeutic agents, such as antibiotics. The pretreatmentwith DMSO, alone or in combination with MSM, enhances the inhalant'stherapeutic effects by at least 10%, 25%, 50%, 100%, 2-fold, 3-fold,5-fold, 10-fold, 50-fold, or 100-fold.

In several embodiments, subjects having drug-resistant tuberculosis aretreated with a formulation comprising, consisting or consistingessentially of DMSO, alone or in combination with MSM, and one or moretherapeutic agents, such as antibiotics. In some embodiments, theformulation additionally includes other therapeutics agents, carriers orexcipients. In one embodiment, the formulation additionally includesarginine, vitamin D, antioxidants, macrolides, linezolid, thioacetazone,thioridazine, or combinations thereof.

Delivery Devices

DMSO readily disrupts the integrity of many materials (particularlyplastics and polymers used in manufacturing disposable medicalequipment). Accordingly, several embodiments of the invention comprisedevices to facilitate the storage and administration of DMSO. In someembodiments, DMSO is stored in glass bottles and administered throughnon-reactive tubing. In other embodiments, inhalant devices arespecially designed to be DMSO resistant. In some embodiments, portionsof the inhalant devices are disposable or replaceable. According toseveral embodiments, formulations comprising DMSO are manufactured,stored and/or administered using materials and devices disclosed in U.S.patent application Ser. No. 12/066,480, which is the National Phaseentry of International Application No.: PCT/US06/35499, filed Sep. 11,2006, which is herein incorporated by reference in its entirety.

In certain embodiments, the delivery device delivers droplets orparticles of the inhaled formulation of a size capable of reaching thebronchioles of the patient's lungs. In some embodiments, activation ofthe delivery device is synchronized with a patient's breathing rhythm tocarry the formulation to the bronchioles. Inhalant therapy according toone embodiment, enables more direct administration the inhaledformulation to infected pulmonary target tissues. Direct targeting isadvantageous in some embodiments because it allows for reduction of theamount of antimicrobial compounds incorporated into the formulationwhile maintaining or improving efficacy of the formulation againstinfectious microorganisms. In other embodiments, direct administrationincreases the efficacy of a given antimicrobial regime against one ormore drug-resistant strains of microorganism. Direct targeting,according to other embodiments, minimizes side effects by minimizingcontact with non-targeted tissue.

The small droplet or particle size that are provided according to someembodiments reduces the volume of DMSO alone or DMSO in combination withMSM that is administered as compared to traditional ventilator therapy.For example, in one embodiment, the use of an inhalant device (e.g.,nebulizer) will be efficacious with about 6 mg to about 25 mg DMSOand/or MSM daily, as compared to 50-100 mg daily when administeredthrough certain other pathways. Reducing DMSO is beneficial in someembodiments because it reduces undesired side effects and odor. In otherembodiments, higher amounts of DMSO are used and tolerated.

In several embodiments, the addition of MSM unexpectedly reduces theunpleasant odor normally experienced with DMSO use. For example, incertain embodiments, DMSO and MSM formulations produce no perceptibleodor after use. In some other embodiments having DMSO concentrationsapproaching or exceeding 50%, the combination with MSM in theformulation reduces or eliminates the DMSO-based odor. Such a result isunexpected, given that DMSO use is normally associated with a strongunpleasant odor.

In some embodiments, the use of DMSO alone or DMSO in combination withMSM with one or more therapeutic agents (such as antibiotics) permitsthe manufacture and/or administration of small droplets or particlesizes, thereby reducing the irritation of the mucosa of the mouth andthroat, as the droplets or particles travel more deeply into the lungsof the patient. In some embodiments, the depth of travel of the dropletsor particles increases the concentration of the dissolved antibiotics inthe patient's lungs.

In several embodiments, DMSO alone or in combination with MSMformulations are combined with therapeutic agents (such as antibiotics)and provided as an aerosol to deliver locally-active drugs to therespiratory system to treat tuberculosis or other respiratory diseases.In one embodiment, the lower airways are contacted (or contactedexclusively) with the formulation. In other embodiments, the formulationis used to systemically treat illnesses. For systemically-active drugs,the aerosol particles are sized to reach the alveolar surface inperipheral areas of the lung.

In some embodiments, the use of DMSO alone or in combination with MSMformulations comprising a therapeutic agent (such as an antibiotic) isparticularly advantageous because it provides rapid onset of action. Inone embodiment, inhalation delivery provides a large absorption area ofthe lung. For locally acting drugs, the onset of action is immediate insome embodiments. Systemically-active inhaled formulations, according tosome embodiments, reach the blood stream quickly. Inhalation therapyprovides a therapeutic effect within about 1-90 minutes in someembodiments, including about 10 to 20 minutes, about 20 to 30 minutes,about 30 to 40 minutes, about 40 to 50 minutes, about 50 to 60 minutes,about 60 to 70 minutes, about 70 to 80 minutes, about 80 to 90 minutes,and overlapping ranges thereof. In one embodiment, DMSO alone or incombination with MSM enhances the bioavailability of the therapeuticagent. In a further embodiment, DMSO alone or in combination with MSMreduces the degradation of the therapeutic agent. In another embodiment,aerosol formulations disclosed herein reduce the gastrointestinal sideeffects or skin irritation that may occur with oral or topicaltreatment.

In several embodiments, inhalant particles are sized to minimize thedeposit of those particles by inertial impact into the upper airwayswithout reaching the site of action. In several embodiments, theparticles are sized to minimize deposit in the mouth and throat, therebyminimizing swallowing and undesired local or systemic side effects. Inseveral embodiments, the particles are smaller than 2, 5 or 10 μm. Inone embodiment, the particles are about 3-5 μm and are transported intothe bifurcations and smaller airways of the bronchii and bronchioles. Inanother embodiment, the particles are less than 3 μm and follow theairflow into the alveoli. In several embodiments, the use of DMSO aloneor in combination with MSM allows for optimizing the particle size ofthe therapeutic agent. Thus, diseases such as tuberculosis can be moreeffectively treated. Moreover, in several embodiments, the use of DMSOalone or in combination with MSM sensitizes drug-resistant tuberculosisto antibiotics and/or renders drug-sensitive microbes more sensitive toan effective therapeutic agent.

In several embodiments, DMSO alone or in combination with MSM forms asolution, mixture, emulsion, suspension, or other suitable combinationwith the therapeutic agent. In one embodiment, homogenization,sonication, high shear fluid processing, or other mechanical methods areused to combine the therapeutic agent with the DMSO or combination ofDMSO and MSM. In other embodiments, the therapeutic agent dissolvesreadily in DMSO. Unlike other strong solvents, DMSO is not harmful tolung tissue. Thus, DMSO is especially advantageous in some embodimentsbecause it can both dissolve the therapeutic agent and deliver saidagent without damaging lung tissue. In some embodiments, DMSO dissolvesat least 50%, 75%, 90%, 95%, or 99% of the therapeutic agent, and in oneembodiment, is able to prevent undesired precipitation of thetherapeutic agent.

In some embodiments, sprays, gels or wipes comprising DMSO, alone or incombination with MSM, and antibacterial agents are provided forsanitizing medical equipment, surfaces and the body to minimize thespread of infectious disease.

Treatment of Drug-Resistant Diseases

In several embodiments, a formulation comprising DMSO alone or incombination with MSM and antimicrobial agents are used as a treatmentfor an infectious disease, including drug-resistant organisms.

In certain embodiments, formulations of DMSO or a combination of DMSOand MSM disclosed herein are effective to treat various infectiousdiseases including, but not limited to, acinetobacter infection,actinomycosis, Adenovirus infection, African sleeping sickness (Africantrypanosomiasis), AIDS, amebiasis, anaplasmosis, Anthrax,Arcanobacterium haemolyticum infection, Argentine hemorrhagic fever,ascariasis, aspergillosis, astrovirus infection, babesiosis, Bacilluscereus infection, bacterial pneumonia, bacterial vaginosis (BV),Bacteroides infection, balantidiasis, Baylisascaris infection, BK virusinfection, black piedra, Blastocystis hominis infection, blastomycosis,Bolivian hemorrhagic fever, Borrelia infection, botulism, Brazilianhemorrhagic fever, brucellosis, Burkholderia infection, Calicivirusinfection, campylobacteriosis, candidiasis (moniliasis; thrush),cat-scratch disease, cellulitis, Chagas disease, chancroid, chickenpox,chlamydia, Chlamydophila pneumoniae infection, cholera,chromoblastomycosis, clonorchiasis, clostridium difficile infection,coccidioidomycosis, Colorado tick fever, common cold, Creutzfeldt-Jacobdisease, Crimean-Congo hemorrhagic fever, cryptococcosis,cryptosporidiosis, cutaneous larva migrans (CLM), cyclosporiasis,cysticercosis, cytomegalovirus infection, dengue fever, dientamoebiasis,diphtheria, diphyllobothriasis, dracunculiasis, Ebola hemorrhagic fever,echinococcosis, ehrlichiosis, enterobiasis (Pinworm infection),Enterococcus infection, enterovirus infection, epidemic typhus, erythemainfectiosum, exanthem subitum, fasciolopsiasis, fasciolosis, fatalfamilial insomnia (FFI), filariasis, food poisoning, free-living amebicinfection, Fusobacterium infection, gas gangrene (Clostridialmyonecrosis), geotrichosis, Gerstmann-Strüssler-Scheinker syndrome(GSS), giardiasis, glanders, gnathostomiasis, gonorrhea, granulomainguinale (Donovanosis), Group A streptococcal infection, Group Bstreptococcal infection, Haemophilus influenzae infection, hand, footand mouth disease (HFMD), Hantavirus, Helicobacter pylori infection,hemolytic-uremic syndrome (HUS), hemorrhagic fever with renal syndrome(HFRS), Hepatitis A, B, C, D, or E, herpes simplex, histoplasmosis,hookworm infection, human bocavirus infection, Hhman ewingiiehrlichiosis, human granulocytic anaplasmosis (HGA), humanmetapneumovirus infection, human monocytic ehrlichiosis, humanpapillomavirus (HPV) infection, human parainfluenza virus infection, andhymenolepiasis.

In certain embodiments, formulations of DMSO or a combination of DMSOand MSM disclosed herein are also effective in treating one or more ofthe following infectious diseases, Epstein-Barr Virus InfectiousMononucleosis (Mono), Influenza (flu), Isosporiasis, Kawasaki disease,Keratitis, Kingella kingae infection, Kuru, Lassa fever, Legionellosis,Leishmaniasis, Leprosy, Leptospirosis, Listeriosis, Lyme disease,Lymphatic filariasis, Lymphocytic choriomeningitis, Malaria, Marburghemorrhagic fever (MHF), Measles, Melioidosis (Whitmore's disease),Meningitis, Meningococcal disease, Metagonimiasis, MicrosporidiosisMicrosporidia, Molluscum contagiosum (MC), Mumps, Murine typhus,Mycoplasma pneumonia, Mycetoma, Myiasis, Neonatal conjunctivitis,Onchocerciasis (River blindness), Paracoccidioidomycosis (South Americanblastomycosis), Paragonimiasis, Pasteurellosis, Pediculosis capitis(Head lice), Pediculosis corporis (Body lice), Pediculosis pubis (Pubiclice, Crab lice), Pelvic inflammatory disease (PID), Pertussis (Whoopingcough), Plague, Pneumococcal infection, Pneumocystis pneumonia (PCP),Pneumonia, Poliomyelitis, Poliovirus, Primary amoebicmeningoencephalitis (PAM), Progressive multifocal leukoencephalopathy,Psittacosis, Q fever, Rabies, Rat-bite fever, Respiratory syncytialvirus, Rhinosporidiosis, Rhinovirus infection, Rickettsial infection,Rickettsialpox, Rift Valley fever (RVF), Rocky mountain spotted fever(RMSF), Rotavirus infection, Rubella, Salmonellosis, SARS (Severe AcuteRespiratory Syndrome), Scabies, Schistosomiasis, Sepsis, Shigellosis,Shingles (Herpes zoster), Smallpox, Sporotrichosis, Staphylococcal foodpoisoning, Staphylococcal infection, Strongyloidiasis, Syphilis,Taeniasis, Tetanus (Lockjaw), Tinea barbae (Barber's itch), Tineacapitis (Ringworm of the Scalp), Tinea corporis (Ringworm of the Body),Tinea cruris (Jock itch), Tinea manuum (Ringworm of the Hand), Tineanigra, Tinea pedis (Athlete's foot), Tinea unguium (Onychomycosis),Tinea versicolor (Pityriasis versicolor), Toxocariasis (Ocular LarvaMigrans (OLM)), Toxocariasis (Visceral Larva Migrans (VLM)),Toxoplasmosis, Trichinellosis, Trichomoniasis, Trichuriasis (Whipworminfection), Tuberculosis, Tularemia, Ureaplasma urealyticum infection,Venezuelan equine encephalitis, Venezuelan hemorrhagic fever, Viralpneumonia, West Nile Fever, White piedra, Yersinia pseudotuberculosisinfection, Yersiniosis, Yellow fever, and Zygomycosis.

In several embodiments, formulations of DMSO or a combination of DMSOand MSM disclosed herein are particularly effective in treating one ormore infectious diseases that are resistant to drug therapies. Inaddition to those infectious diseases listed above, which may already beor may become drug resistant in the future, certain embodiments usingDMSO or a combination of DMSO and MSM are effective in treating, amongothers, drug resistant: tuberculosis, measles, tetanus, malaria, upperand lower respiratory infections, hepatitis, typhoid fever,vancomycin/glycopeptide-intermediate Staphylococcus aureus infection,vancomycin-resistant enterococci, methicillin-resistant Staphylococcusaureus (MRSA), and streptococcus pneumoniae.

In some embodiments, treatment of an infectious disease comprises thepretreatment of a patient with DMSO, followed by the administration of aformulation comprising DMSO and antimicrobial agents. In otherembodiments, treatment of an infectious disease comprises thepretreatment of a patient with DMSO, followed by the administration of aformulation comprising DMSO, MSM, and antimicrobial agents. In someembodiments, DMSO pretreatment is administered intravenously via a fastdrip IV catheter. In other embodiments, the DMSO is given in a bolus IVinjection. In yet other embodiments, pretreatment with DMSO is notperformed. Pretreatment compositions additionally include MSM, atherapeutic agent or a combination thereof in some embodiments.

In several embodiments, formulations comprising DMSO and antimicrobialagents, or DMSO, MSM and antimicrobial agents are administered orally,intravenously, intramuscularly, or subcutaneously. However, as the siteof infection of several infectious diseases, including tuberculosis, isthe lungs, in some embodiments, formulations are administered byinhalation. In some such embodiments, the inhalant means comprises anebulizer. In other embodiments, an inhaler is used.

As discussed above, the use of a DMSO alone or in combination with MSMformulation comprising an antimicrobial formulation in a nebulizer or aninhaler provides particular advantages in several embodiments. Incertain embodiments, the dose of antimicrobial compounds that reduce oreliminate microorganisms is reduced compared to using the sameantimicrobial compounds alone. In other embodiments, the delivery of aformulation as described herein, via a nebulizer or inhaler, is moreeffective at reducing or eliminating infectious microorganisms ascompared to intravenous, intramuscular, oral, or standard ventilatorbased therapy. In addition, the use of nebulizers or inhalers in someembodiments provides the improved ability to bring treatment to patientsin geographical areas that are not amenable to standard treatmenttechniques requiring hospitalization time. The efficacious therapy andimproved portability realized with some embodiments is especiallybeneficial given the long-felt, but unmet need for such a type oftherapy.

In several embodiments, subjects are pretreated with DMSO usingintravenous DMSO by fast drip within, e.g., a ten minute period (orless). In one embodiment, DMSO will be provided in glass bottles withproprietary non-reactive tubing. Subjects will then receive antibioticsdissolved in DMSO in 3 ml doses through an inhaler or mouth spray threetimes a day with meals. In one embodiment, DMSO pretreatment is providedin the range of about 25 mg to about 75 mg (e.g., 30 mg, 40 mg, 50 mg,60 mg, 70 mg) in 200 ml 5% dextrose and water. In one embodiment, 56 mgDMSO in 200 ml 5% dextrose and water is provided. In one embodiment, thefollowing antibiotics are provided: rifampicin, isoniazid, pyrazinamide,and ethambutol. In one embodiment, about 600 mg rifampicin, 300 mgisoniazid, 2.4 g pyrazinamide, and 1.2 g ethambutol are administered perday, through an inhaler/nebulizer or mouth spray delivered in 3 mldosages, three times daily. In one embodiment, the antibiotics arecombined with DMSO for delivery via inhalation, with or without thepretreatment with DMSO. Pretreatment with MSM is also provided inseveral embodiments. Intravenous pretreatment of DMSO, or a combinationof DMSO and MSM is provided in some embodiments. Pretreatmentformulation may also comprises therapeutic agents.

In several embodiments, therapeutic effects are obtained within twoweeks of treatment, within two months of treatment, and/or within sixmonths of treatments. Other therapeutic windows are also provided.

In several embodiments, subjects who test positive for the presence ofthe M. tuberculosis complex and are resistant one or more antibiotics(such as rifampicin and isoniazid) are identified and treated. Aftertreatment with a formulation comprising DMSO alone or in combinationwith MSM and rifampicin, isoniazid, pyrazinamide, and ethambutol, thetreated subjects will show a negative sputum mycobacterial cultureand/or show improvement in one or more of the following symptoms:coughing, chest pain, shortness of breath, labored breathing, weightloss, fatigue, fever, night sweats, chills, and loss of appetite. Inother embodiments, treated subjects will show a greater reduction in M.tuberculosis colonies resulting from the culturing of sputum samples ascompared to control group patients who are not provided DMSO alone or acombination of DMSO and MSM. In some embodiments, in addition to, or ininstead of, reducing the number of M. tuberculosis colonies culturedfrom sputum samples, the overall health and robustness of the resultingcolonies is reduced, indicating potentially reduced severity ofinfection as well as demonstrating that the DMSO-based treatment isnegatively impacting the M. tuberculosis.

In some embodiments, patients pretreated with DMSO show betterimprovement than those treated with inhalant DMSO and antibioticswithout intravenous DMSO pretreatment. In some embodiments, patientstreated with DMSO with inhalant DMSO and antibiotics show betterimprovement than those treated with antibiotics alone. In severalembodiments, the addition of MSM to the formulation containing DMSOenhances the therapeutic effects or reduces side effects. In oneembodiment, MSM is used alone as a pretreatment.

In several embodiments, formulations for treating malaria are provided.In one embodiment, a formulation comprising about 20%-60% (e.g. 40%)DMSO, about 5%-20% (e.g., 10%) MSM and, optionally, about 10%-40% (e.g.,20%) urea. In one embodiment, about two hundred milliliters (200 ml) areadministered intravenously to malaria patients. Other amounts may alsobe administered. In some embodiments, an orally administered herb (e.g.,qinghao, also known as sweet wormwood or Artemisia annua) is alsoadministered. This treatment regime is administered a single time (ormay be administered multiple times). Patients show improvements inmalarial symptoms after 24 hours (e.g., reduced fever, vomiting, reduceddiarrhea). In addition to the reduction in malarial symptoms, theunpleasant odor normally associated with administration of DMSO issignificantly reduced.

In several embodiments, DMSO, alone or in combination with MSM, andoptionally in combination with urea advantageously reduces symptoms ofmalaria. In some embodiments, the inclusion of a traditional medicine(such as an herb or an antibiotic) further enhances the efficacy of theformulation. As discussed above, in several embodiments DMSO, alone orin combination with MSM, functions to sensitize bacteria to atherapeutic agent. In several embodiments, subjects treated with a DMSOalone or in combination with MSM formulation include humans, non-humanmammals, domesticated animals and livestock. In some embodiments,formulations disclosed herein are used to not only treat undesiredsymptoms and illnesses, but can also act as a preventative agent. Forexample, formulation may be taken on a regular basis to prevent theonset of illness (e.g., tuberculosis). In one embodiment, at risksubjects (e.g., family members or subjects who are exposed to patientshaving tuberculosis) will be administered lower doses of DMSO alone orin combination with MSM and antibiotics to prevent the onset ofinfection.

EXAMPLES

As discussed herein, MSM is used in several embodiments in conjunctionwith one or more antibiotics and/or DMSO in order to treat infectiousdiseases. The following studies were directed to determining theantimicrobial properties of both MSM and DMSO, either alone or incombination, as well as their efficacy when used in conjunction withpenicillin.

The microorganisms used to test MSM, DMSO, penicillin and theircombinations were Streptococcus pyogenes and Streptococcus pneumonia,based on these two organisms being responsible for a variety of humanstreptococcal infections. S. pyogenes has a hyaluronic acid capsulewhile S. pneumonia has a distinct polysaccharide capsule. The distinctcarbohydrate composition of bacterial antigens found on the cell wallsof these two microorganisms led S. pyogenes to be classified asLancefield Group A and S. pneumonia to remain unclassified underLancefield Group standards. The use of related, but structurallydistinct microorganisms was intended to assist in determining if MSM,DMSO, penicillin and their combinations were more effective on one typeof bacteria over the other.

Thus, the purpose of these studies was to determine 1) the antimicrobialeffects of DMSO, both individually and in combination with MSM, on S.pyogenes and S. pneumonia; 2) the most effective concentrations forantimicrobial properties for such; 3) whether combining MSM and DMSOreduces the concentrations of either compound needed to achievetherapeutically relevant levels of microbial reduction; and 4) theantimicrobial effectiveness of using DMSO, and the combination of DMSOAND MSM in conjunction with an antibiotic agent.

S. pneumonia (#10341™) and S. pyogenes (#10096™) were purchased fromATCC. DMSO (#D8418) was purchased from Sigma-Aldrich. Penicillin waspurchased from Henry Schein. Bacterial culture medium was purchased fromBecton-Dickinson. A bioluminescent adenosine triphosphate (ATP) assaykit was purchased from Promega (#G8230).

S. pyogenes and S. pneumonia were cultured in Brain Heart Infusion media(BD 237500) at 37° C. Aliquots of the cultures were plated and exposedto the appropriate concentrations of DMSO, MSM, and/or antibiotic.Bacteria were cultured for 7 hours (for Streptococcus pneumonia) and 18hours (for Streptococcus pyogenes) respectively. Then bacterialviability was evaluated by the Promega bioluminescent ATP assay kit. Thetest was conducted in triplicate.

Analysis of bacterial ATP content was used to evaluate cell viability.Bacterial ATP can be measures by direct lysis of the bacteria with adetergent, thereby releasing ATP from the cells. The released ATP reactswith user supplied luciferase and results in light emission. Theintensity of the emitted light is measured with a luminometer and isproportional to the concentration of ATP. Greater concentrations of ATPare indicative of greater bacterial viability. Briefly, the reagentswere prepared according the to the manufacturer's instructions. Forexample, opaque multiwall tubes or microplates (depending on the numberof samples to be evaluated) are used to culture bacteria as discussedherein. Control tubes (culture medium without cells) are prepared togenerate a value for background luminescence. Luciferase containingreagent (BacTiterGlo™) is added to each tube/well (including controltubes). After mixing thoroughly and incubating the mixture forapproximately 5 minutes at room temperature, luminescence is measuredusing a luminometer.

For S. pneumonia, MSM and DMSO were prepared in concentrations of 5%,10%, and 20%. Penicillin was prepared at concentrations of 25, 50, and100 μg/L. For S. pyogenes, MSM was prepared in concentrations of 0.31,0.63, 1.25, 2.50, 5.00, 10.0, 20.0 and tested for synergy atconcentrations of 2.5, 5, and 10%; DMSO was prepared in concentrationsof 0.31, 0.63, 1.25, 2.50, 5.00, 10.0, 20.0, and tested for synergy atconcentrations of 2.5, 5, and 8%; and penicillin was prepared inconcentrations 1.56, 3.13, 6.25, 12.5, 25.0, 50, 100 μg/L and tested forsynergy at concentrations of 3.125, 6.25, and 12.5 μg/L. MSM, DMSO andpenicillin were all diluted in culture medium. The synergistic effectsof the various compounds in combination were then evaluated. As usedherein, the term synergistic shall be given its ordinary meaning andshall also refer to instances wherein the combination of two or moresubstances kills a higher percentage of a microorganism than would beanticipated by adding the individual kill rates. Synergistic effects areindicated by an “*” in the Tables that follow.

Example 1 Antibacterial Effects of DMSO and MSM Alone on S. pneumonia

DMSO was added to S. pneumonia cultures to final concentrations of 5%,10%, and 20%. At each of these concentrations, DMSO resulted inreductions in bacterial viability in a dose-dependent manner. SeeTable 1. Thus, in some embodiments, DMSO alone, at concentrationsbetween 5 and 20% is effective at reducing the viability of certainbacteria. In several embodiments, DMSO also provides analgesic and/oranti-inflammatory effects, which may be beneficial, as pain and/orinflammation are associated with certain bacterial infections.

TABLE 1 Viability of S. pneumonia After DMSO Exposure DMSO ConcentrationViability of S. pneumonia (%) 5 75.45 10 40.18 20 27.19

MSM alone was added to S. pneumonia cultures to final concentrations of5%, 10%, and 20%. At each of these concentrations, MSM also resulted inreductions in bacterial viability in a dose-dependent manner. See Table2. The extent of reduction in bacterial viability was not as robust withMSM as compared to DMSO, however, in several embodiments, MSM is stilluseful as an antimicrobial agent when used alone. In severalembodiments, MSM alone is preferable as an antimicrobial agent becauseof its beneficial analgesic and anti-inflammatory properties, as bothpain and inflammation may be associated with certain infections.

TABLE 2 Viability of S. pneumonia After MSM Exposure MSM ConcentrationViability of S. pneumonia (%) 5 95.34 10 48.57 20 39.08

Example 2 Antibacterial Effects of Combinations of DMSO and MSM on S.pneumonia

In accordance with several embodiments described above, MSM and DMSO incombination were evaluated for their antibacterial effects on S.pneumonia. DMSO at 5%, 10%, and 20% was combined with MSM at 0% (DMSOonly control), 5%, 10%, or 20%. As shown in Tables 3, 4, and 5, certaincombinations of MSM with DMSO are synergistic as compared to either DMSOor MSM alone. Synergistic results as compared to DMSO or MSM alone areindicated by an “*”. For example, addition of 5% MSM to 5% DMSO reducedbacterial viability to approximately 50% (see Table 3), while thecombined effects of 5% MSM and 5% DMSO would be expected to be areduction in bacterial viability to approximately 70%. Similarly, 5% MSMadded to 20% DMSO reduced bacterial viability to approximately 18% (seeTable 5), while combined effects of 5% MSM and 20% DMSO would beexpected to be a reduction in bacterial viability to only about 22%. Incontrast, 5% MSM in combination with 10% DMSO did not result insynergistic reductions in bacterial viability. Thus, the interaction of5% MSM with DMSO exhibits a biphasic effect curve, with 5% and 20% DMSOin combination with 5% MSM resulting in unexpectedly enhancedanti-microbial effects.

These results therefore indicate that certain combinations of DMSO andMSM are more effective as antibacterial agents. Thus, in severalembodiments, about 5% MSM is used in conjunction with about 5% DMSO toyield unexpected reductions in bacterial viability. In some embodiments,about 5% MSM is used in conjunction with about 20% DMSO to yieldunexpected reductions in bacterial viability. Certain such embodimentsare particularly advantageous because both DMSO and MSM have analgesicand anti-inflammatory properties, both of which may be beneficial inreducing symptoms of certain microbial infections. Moreover, in someembodiments employing a larger (e.g., 20%) concentration of DMSO, theMSM in the formulation functions to reduce one or more of the sideeffects of DMSO (e.g., odor associated with DMSO administration). Instill additional embodiments, those combinations that show synergy inreducing microbial viability also reduce the amount of time for thecompounds to affect viability. In some embodiments, such effects reducethe overall treatment time (e.g., complete resolution of an infection)and/or reduce the time to demonstrable improvements in symptoms (e.g.,reduced time to apyresis). In still additional embodiments, thosecombinations of DMSO and MSM that did not demonstrate synergisticeffects on bacterial viability may still beneficially (andsynergistically) impact treatment times. In some embodiments, asdiscussed below, certain combinations of MSM and DMSO render anotheragent more efficacious (e.g., same effects at a lower administereddose).

TABLE 3 Viability of S. pneumonia After Exposure to VariousConcentrations of MSM in 5% DMSO DMSO (%) MSM (%) S. pneumonia viability(%) 5 0 75.45 5 5 50.94* 5 10 45.40 5 20 27.22

TABLE 4 Viability of S. pneumonia After Exposure to VariousConcentrations of MSM in 10% DMSO DMSO (%) MSM (%) S. pneumoniaviability (%) 10 0 40.18 10 5 47.81 10 10 37.42 10 20 11.95

TABLE 5 Viability of S. pneumonia After Exposure to VariousConcentrations of MSM in 20% DMSO DMSO (%) MSM (%) S. pneumoniaviability (%) 20 0 27.19 20 5 17.60* 20 10 7.76 20 20 5.15

Example 3 Antibacterial Effects of DMSO Alone or in Combination with MSMin Further Combination with Penicillin on S. pneumonia

As discussed herein, in several embodiments, the addition of DMSO aloneor in combination with MSM to a therapeutic agent, such as anantibiotic, acts to reduce the effective concentration of thetherapeutic agent. For example, in some embodiments, DMSO alone or incombination with MSM in combination with an antibiotic reduces theamount of antibiotic needed to achieve reductions in the viability oftarget bacteria. In some embodiments, the amount of penicillin, oranother beta-lactam antibiotic needed for efficacious reduction inbacterial viability is reduced. In other embodiments, other antibioticswith other mechanisms of action (e.g., aminoglycosides, tetracyclines,macrolides, among others) are needed in reduced concentrations. In someembodiments, the reduction of the amount of antibiotic (or othertherapeutic agent) needed for an effect also reduces one or more sideeffects associated with the agent. For example, some antibiotic agentsare associated with gastric discomfort and/or diarrhea, due to thereduced viability of native gastrointestinal flora. Other antibioticshave side effects such as reduced appetite, dry mouth, and the like. Insome instances, the route of delivery, as discussed above, can be usedto partially manage the side effects. However, in some embodiments,reduction in the amount of drug needed for efficacious treatment is moreadvantageous in reducing side effects. Additionally, in someembodiments, the reduction in the amount of antibiotic agent required toreduce microbial viability reduces the incidence of allergic eventsinduced by the antibiotic. Moreover, the reduction in amount of agentneeded for therapy, in some embodiments, increases the number of doses(and therefore possibly the number of patients) that may be treated witha given amount of agent.

DMSO in Combination with Penicillin

DMSO, at 0, 5, 10, or 20% was individually combined with 25, 50, or 100μg/L penicillin and the combinations were evaluated for their effects onbacterial viability. As shown in Tables 6-8, no synergistic reductionsin bacterial viability were detected with any combinations of varyingDMSO concentrations and varying penicillin concentrations. As discussedabove however, these combinations, in some embodiments, may still beeffective at reducing the treatment time (either to resolution or toreduction of symptoms). These data do indicate, however, that certaincombinations of DMSO and penicillin result in equally efficaciousreductions in bacterial viability, with a decreased amount of penicillin(e.g., DMSO functioning to reduce the effective concentration ofpenicillin).

For example, 20% DMSO in combination with 100 μg/L penicillin yieldedapproximately 80% reduction in bacterial viability (21.76% viability,see Table 8). 20% DMSO in combination with 50 μg/L penicillin alsoreduced bacterial viability by about 80% (19.14% viability, see Table7). Finally, 20% DMSO in combination with only 25 μg/L penicillin stillreduced bacterial viability by 78% (22.08% viability, see Table 6).Thus, when combined with 20% DMSO, the effective concentration ofpenicillin can be reduced by 5-fold, while still achieving the samereduction in bacterial viability. As discussed above, this type ofsynergy is particularly advantageous in some embodiments, as theincidence of allergic reactions to an antibiotic agent may be reduceddue to the lower concentration of antibiotic needed. Moreover, in somesuch embodiments, the rate of generation of microbial resistance may bereduced, again due to the reduction in the necessary amount ofantibiotic. In some embodiments, the reduced amount of antibiotic alsoreduces the incidence of antibiotic related side effects. In stilladditional embodiments, despite the reduction in the amount ofantibiotic required to reach a given therapeutic effect, the time toreach that therapeutic effect is reduced (e.g., treatment time reducedfrom weeks to days). Additionally, a given amount of penicillin may beused to treat a greater number of patients (or a single patient agreater number of times). This may be of particular importance in remotelocations, where transport and/or storage of a drug is more difficult

In some embodiments, the penetrant nature of DMSO functions to increasethe amount of penicillin able to enter the bacterial wall and inhibitformation of peptidoglycan cross-links. Likewise, in other embodiments,DMSO allows other antibacterial or therapeutic agents (e.g., nonbeta-lactam antibiotics) greater access to the interior of the bacteria,enhancing the efficacy of the antibiotic. In some embodiments, the DMSOprovides favorable effects for a patient, such as, pain reduction,anti-inflammatory effects, and the like.

Similar trends are present in the when 10% and 5% DMSO is combined withpenicillin, in that similar reductions in bacterial efficiency areachieved, despite using lower concentrations of penicillin (see e.g.,10% and 5% data points in Tables 6-8). In some embodiments, reducedcomparative efficacy of the 10% or 5% DMSO/penicillin combinations isnot of clinical significance (e.g., for treatment of a low grade,non-emergency infection). As such, despite the lesser overall reductionin bacterial viability, combinations of 10% or 5% DMSO with penicillinare preferred in some embodiments. In such embodiments, the reducedamount of DMSO may be advantageous, in that DMSO-associated side effectsmay be reduced (e.g., skin or mucous membrane irritation; odorassociated with administration).

TABLE 6 S. pneumonia Viability After Exposure to Various Concentrationsof Penicillin Penicillin (μg/L) S. pneumonia viability (%) 25 79.82 5042.70 100 40.93

TABLE 7 S. pneumonia Viability After Exposure to 25 μg/L of Penicillinwith Various Concentrations of DMSO Penicillin (μg/L) DMSO (%) S.pneumonia viability (%) 25 0 79.82 25 5 46.13 25 10 39.78 25 20 22.08

TABLE 8 S. pneumonia Viability After Exposure to 50 μg/L of Penicillinwith Various Concentrations of DMSO Penicillin (μg/L) DMSO (%) S.pneumonia viability (%) 50 0 42.70 50 5 46.27 50 10 37.09 50 20 19.14

TABLE 9 S. pneumonia Viability After Exposure to 100 μg/L of Penicillinwith Various Concentrations of DMSO Penicillin (μg/L) DMSO (%) S.pneumonia viability (%) 100 0 40.93 100 5 45.09 100 10 35.80 100 2021.76MSM in Combination with Penicillin

The combination of 5% MSM with 25 μg/L of penicillin exhibited asynergistic reduction in the viability of S. pneumonia, leading to only41% viability (see Table 10). Synergy as compared to the expectedresults based on MSM alone and penicillin alone is indicated in theTables by an “*”. In contrast, 5% MSM alone reduced viability by only˜5%, while 25 μg/L penicillin alone reduced viability by ˜21%. Thus, the5% MSM/25 μg/L of penicillin combination is unexpectedly moreefficacious than expected based on the results obtained with MSM orpenicillin alone.

Moreover, as with DMSO, certain concentrations of MSM allowed lowerconcentrations of penicillin to reduce bacterial viability nearly aseffectively as higher concentrations. For example, 20% MSM with 100 μg/Lpenicillin reduced S. pneumonia viability to 21.37%, (see Table 12) 20%MSM with 50 μg/L penicillin reduced S. pneumonia viability to 20.75%(see Table 11). Thus, with use of 20% MSM, the required concentration ofpenicillin is reduced by one-half. Continuing this trend is thecombination of 20% MSM with 25 μg/L penicillin reduced S. pneumoniaviability to approximately 25% (see Table 10). Thus, in someembodiments, if antibiotic related side effects are of concern, certainconcentrations of MSM are unexpectedly advantageous in allowing reducedlevels of antibiotic to be administered. Similarly, though with a lessrobust reduction in bacterial viability, 5% MSM allowed 25 μg/Lpenicillin to perform nearly identically to 100 μg/L penicillin (compareTables 10-12 for 25 μg/L penicillin).

TABLE 10 S. pneumonia Viability After Exposure to 25 μg/L of Penicillinwith Various Concentrations of MSM Penicillin (μg/L) MSM (%) S.pneumonia viability (%) 25 0 79.82 25 5 41.23* 25 10 41.83 25 20 25.36

TABLE 11 S. pneumonia Viability After Exposure to 50 μg/L of Penicillinwith Various Concentrations of MSM Penicillin (μg/L) MSM (%) S.pneumonia viability (%) 50 0 42.70 50 5 41.23 50 10 47.47 50 20 20.75

TABLE 12 S. pneumonia Viability After Exposure to 100 μg/L of Penicillinwith Various Concentrations of MSM Penicillin (μg/L) MSM (%) S.pneumonia viability 100 0 40.93 100 5 41.75 100 10 36.67 100 20 21.37

Example 4 Antibacterial Effects of DMSO and MSM in Combination withPenicillin on S. pneumonia

Based on the synergistic results seen in certain combinations of MSM orDMSO with penicillin, the present experiment was performed in order toidentify the various combinations of MSM, DMSO, and penicillin thatyielded synergistic reductions in bacterial viability as compared tocombination the effects of combining DMSO, MSM, and penicillin onbacterial viability. This experiment was also designed to identifycombinations of the three compounds that advantageously allow one ormore of the compounds to be reduced, yet still efficaciously reducebacterial viability.

DMSO at 5, 10, and 20% was combined individually with MSM at one of 5,10, or 20% and penicillin at one of 25, 50, or 100 μg/L. Viability wasassessed as described above. Viability data are presented in Table 13.The “*” symbol represents synergistic results as compared to thecorresponding combination of DMSO and penicillin. The “ψ” symbolrepresents synergistic results as compared to the correspondingcombination of MSM and penicillin. The values for the reduction inbacterial viability were added together to determine the thresholdreduction for synergy. For example, 5% DMSO reduces viability byapproximately 25% and 25 μg/L penicillin reduced viability byapproximately 21%, for a total combined reduction expected ofapproximately 46%. This represents a 64% viability. Thus, if thecombination of 5% MSM, 5% DMSO, and 25 μg/L penicillin results in lessthan 64% viability, synergy between the compounds has been identified.

Several combinations of MSM, DMSO, and penicillin yield synergisticimprovements in bacterial reduction. For example, the combination of 5%DMSO, 5% MSM, and 25 μg/L penicillin reduced bacterial viability toapproximately 52% (see Table 13). 5% DMSO in combination with 25 μg/Lpenicillin reduced bacteria viability to approximately 64% (e.g., abouta 46% reduction, based on the individual reduction seen with 5% DMSO,see Table 1, and the individual reduction seen with 25 μg/L penicillin,see Table 5). Thus the combination of all three compounds reducedbacterial viability by about an additional 12%. Similarly, thecombination of 5% MSM with 25 μg/L penicillin resulted in bacteriaviability of about 74%, while the combination of all three compoundsreduced viability by nearly an additional 22%.

In some combinations, synergistic results were detected with respect toboth DMSO and penicillin as well as MSM plus penicillin. For example,10% DMSO in combination with 20% MSM and 25 μg/L penicillin is yielded asynergistic improved in antimicrobial activity as compared to bothreference combinations. Thus in some embodiments, the combination of allthree compounds outperforms DMSO plus penicillin and MSM pluspenicillin. In some embodiments, such combinations are particularlyadvantageous in combating infections, due to their unexpected efficacy.As such, those combinations, in some embodiments, result in a morerapid, more complete, or more effective reduction and/or elimination ofan unwanted bacterial infection.

In other combinations, synergy was detected only with respect to eitherDMSO plus penicillin or MSM plus penicillin. For example, thecombination of 5% MSM with 10% DMSO and 25 μg/L penicillin wassynergistic with respect to MSM plus penicillin, but not with respect toDMSO plus penicillin. In some embodiments, such information is useful,for example if an individual is sensitive to one of the three compounds,it may be that the offending compound can be removed from thecombination, but effective treatment can still result with only twocompounds. In other embodiments, the differential synergy is due to thehighly effective nature of one or more of the compounds when used alone.As a result, synergy may be mathematically impossible. As discussedabove, in some such embodiments, the combination of all three compoundsincreases rate of reduction in bacterial viability and increases theduration of reduction (e.g., prevents regrowth and/or reinfection).Thus, even when a synergistic reduction in the overall viability ofbacteria is not recognizable, in certain embodiments, other synergisticresults are still detectable.

In addition to the synergistic effects discussed above, there areseveral instances wherein the certain combinations of DMSO, MSM andpenicillin allow for a reduction in the efficacious concentration ofpenicillin. For example, as shown in Table 13, the combination of 5%DMSO with 20% MSM yields very similar overall bacterial viability overthe range of penicillin concentrations tested (from ˜25% viability with25 μg/L penicillin to ˜18% viability with 100 μg/L penicillin).Additionally, 10% DMSO with 20% MSM resulted in nearly identicalbacterial viabilities across the penicillin concentration range.

Similar results are seen with 20% DMSO in combination with 5, 10, or 20%MSM and any concentration of penicillin. These results reveal a slightlywider range of bacterial viability across the different penicillinconcentrations, however, given that the reduction in all casesapproaches approximately 90 to 95%, these combinations are all stillvery effective.

In some embodiments, therefore, certain combinations of DMSO and MSMallow for the reduction in penicillin concentrations. In someembodiments, the presence of 20% MSM allows penicillin to be reducedwhile significant bacterial reductions are realized. In otherembodiments, the presence of 20% DMSO allows penicillin to be reducedwhile significant bacterial reductions are realized. In still additionalembodiments, the presence of 20% MSM and 20% DMSO allows penicillin tobe reduced while significant bacterial reductions are realized. Theseembodiments are particularly advantages in avoiding complications due toadverse reactions to penicillin (e.g., allergies, side effects, etc.)Thus in some embodiments, methods for reducing side-effects or adversereactions to an antibiotic are provided. In some embodiments, 20% MSM,20% DMSO, or 20% MSM and 20% DMSO reduce the effective concentration ofantibiotic required to generate a therapeutic reduction in bacterialviability. In still additional embodiments, other aspects of reducingbacterial viability are maintained (and/or improved) even with areduction in the amount of penicillin (e.g., the rate of reduction inbacterial viability, the duration of reduction, etc.).

TABLE 13 S. pneumonia Viability After Exposure to Various combinationsof DMSO, MSM, and Penicillin DMSO (%) MSM (%) Penicillin (μg/L) S.pneumonia viability (%) 5 5 25 52.11*,^(ψ) 5 5 50 43.36 5 5 100 53.03 510 25 51.82* 5 10 50 44.52 5 10 100 31.33 5 20 25 24.91* 5 20 50 19.20 520 100 18.12 10 5 25 44.41^(ψ) 10 5 50 38.24 10 5 100 36.19 10 10 2539.38 10 10 50 33.73 10 10 100 25.98 10 20 25 11.87*,^(ψ) 10 20 50 11.0310 20 100 10.96 20 5 25 12.74*,^(ψ) 20 5 50 13.39^(ψ) 20 5 100 9.28^(ψ)20 10 25 7.69*,^(ψ) 20 10 50 7.74 20 10 100 5.58 20 20 25 4.93*,^(ψ) 2020 50 5.60 20 20 100 1.80

Example 5 Antibacterial Effects of DMSO and MSM Alone on S. pyogenes

As discussed above, the structure of S. pyogenes differs from that of S.pneumonia, and therefore additional experiments were undertaken toevaluate the synergistic effects of various concentrations of DMSO andMSM, as well as combinations of DMSO, MSM, and penicillin.

DMSO was added to S. pyogenes cultures to final concentrations of 0.31,0.63, 1.25, 2.50, 5.00, 10.0, or 20.0%. At these concentrations, DMSOresulted in reductions in bacterial viability in a dose-dependentmanner. See Table 14. Thus, in some embodiments, DMSO alone, atconcentrations between 5 and 20% is effective at reducing the viabilityof certain bacteria. In several embodiments, DMSO also providesanalgesic and/or anti-inflammatory effects, which may be beneficial, aspain and/or inflammation are associated with certain bacterialinfections.

MSM alone was added to S. pyogenes cultures to final concentrations of0.31, 0.63, 1.25, 2.50, 5.00, 10.0, or 20.0%. At these concentrations,MSM also resulted in reductions in bacterial viability in adose-dependent manner. See Table 15. Thus, in some embodiments, MSMalone, at concentrations between 5 and 20% is effective at reducing theviability of certain bacteria. In several embodiments, MSM also providesanalgesic and/or anti-inflammatory effects, which may be beneficial, aspain and/or inflammation are associated with certain bacterialinfections.

TABLE 14 Viability of S. pyogenes After DMSO Exposure DMSO ConcentrationViability of S. pyogenes (%) 0.31 100 0.63 100 1.25 100 2.50 100 5.0096.66 10.0 14.50 20.0 5.14

TABLE 15 Viability of S. pyogenes After MSM Exposure MSM ConcentrationViability of S. pyogenes (%)) 0.31 100 0.63 100 1.25 100 2.50 100 5.0095.88 10.0 23.94 20.0 15.18

Example 6 Antibacterial Effects of DMSO and MSM in Combination on S.pyogenes

In accordance with several embodiments described above, MSM and DMSO incombination were evaluated for their antibacterial effects on S.pyogenes. DMSO at 2.5%, 5%, and 8% was combined with MSM at 0% (DMSOonly control), 2.5%, 5%, and 10%. As shown in Tables 16, 17, and 18certain combinations of MSM with DMSO are synergistic as compared to theeffects of either DMSO or MSM alone. Synergistic results as compared toDMSO or MSM alone are indicated by an “*”. For example, addition of 2.5%MSM to 2.5% DMSO reduced bacterial viability to approximately 65% (seeTable 16), while the no effect of these concentrations of MSM and DMSOwould be expected, as individually, neither compound reduced bacterialviability. The synergistic effect is also seen with 2.5% DMSO and 5%MSM, where bacterial viability is reduced by nearly 83% (as compared toan expected 4% reduction based on the compounds' effects alone). Synergyis also seen with 5% DMSO in combination with any concentration of MSM.Thus, in some embodiments, DMSO at 5% induces synergistic reductions inbacterial viability in combination with any concentration of MSM between2.5% and 10%. In some embodiments, DMSO at 2.5% and MSM inconcentrations between 2.5% and 5% are advantageously and unexpectedlysynergistic at reducing bacteria viability.

TABLE 16 Viability of S. pyogenes After Exposure to VariousConcentrations of MSM in 2.5 DMSO DMSO (%) MSM (%) S. pyogenes viability(%) 2.5 0 100 2.5 2.5 65.06* 2.5 5.0 17.71* 2.5 10.0 16.37

TABLE 17 Viability of S. pyogenes After Exposure to VariousConcentrations of MSM in 5 DMSO DMSO (%) MSM (%) S. pyogenes viability(%) 5.0 0 96.66 5.0 2.5 36.21* 5.0 5.0 7.87* 5.0 10.0 7.64*

TABLE 18 Viability of S. pyogenes After Exposure to VariousConcentrations of MSM in 8 DMSO DMSO (%) MSM (%) S. pyogenes viability(%) 8.0 0 9.96 8.0 2.5 14.37 8.0 5.0 5.97 8.0 10.0 5.60

Example 7 Antibacterial Effects of DMSO or MSM in Combination withPenicillin on S. pyogenes

Various concentrations of penicillin alone were evaluated for theirability to reduce viability of S. pyogenes. As shown in Table 19,penicillin decreased bacterial viability in a dose-dependent fashion.

TABLE 19 S pyogenes Viability After Exposure to Various Concentrationsof Penicillin Penicillin (μg/L) S. pyogenes viability (%) 1.56 100 3.13100 6.25 100 12.5 13.16 25.0 9.07 50 9.57 100 9.40DMSO in Combination with Penicillin

Due to the highly efficacious nature of penicillin concentrations at orabove 25 μg/L, DMSO was combined with concentrations of penicillin thatwere less efficacious (ranging from 3.125 to 12.5 μg/L). As such,identification of synergism between DMSO and penicillin would be lesslikely to be mathematically obscured.

As shown in Tables 20, 21, and 22 (identified by an “*”) severalcombinations of DMSO and penicillin resulted in synergistic results. Forexample, 5% DMSO in combination with 3.125 μg/L penicillin, based on theefficacy of the two compounds alone, would only be expected to reducebacteria viability by about 4%. However, when combined, the actualreduction was approximately 10-fold greater (viability reduced to ˜61%,see Table 20). Similar synergistic effects were seen when 5% DMSO wascombined with 6.25 μg/L or 12.5 μg/L penicillin (see Table 21 and 22,respectively). Thus, in several embodiments, combinations of DMSO andpenicillin that, taken alone are ineffective, unexpectedly reducebacterial viability. Such embodiments are particularly advantageous inthat they may allow for treatment of a patient that was thought to berefractory to the individual compounds. Moreover, substantial synergy isdetected after combining relatively low concentrations of each of thetwo compounds. As such, side-effects may be minimized.

In the event that side effects are at issue, certain embodiments allowthe reduction of one or more of the compounds while still retaining arelatively efficacious effect on bacterial viability. For example, ifside-effects from DMSO are experienced, a concentration of 12.5 μg/Lpenicillin in combination with 5% DMSO (a 3% reduction in DMSO) stillreduces bacterial viability to approximately 15% of the startingpopulation. Similarly, if penicillin presents issues, the combination of8% DMSO with 6.25 μg/L penicillin is virtually indistinguishable from 8%DMSO with 12.5 μg/L penicillin. Thus, penicillin concentrations can bereduced by 50%, while efficacy is retained. Similar results occur with5% DMSO and 6.25 μg/L penicillin. Thus, in some embodiments, there areprovided methods and compositions for reducing the efficaciousconcentration of DMSO and/or penicillin.

TABLE 20 S pyogenes Viability After Exposure to 3.13 μg/L of Penicillinwith Various Concentrations of DMSO Penicillin (μg/L) DMSO (%) S.pyogenes viability (%) 3.13 0 100 3.13 2.5 100 3.13 5.0 60.85* 3.13 8.012.90

TABLE 21 S pyogenes Viability After Exposure to 6.25 μg/L of Penicillinwith Various Concentrations of DMSO Penicillin (μg/L) DMSO (%) S.pyogenes viability (%) 6.25 0 100 6.25 2.5 100 6.25 5.0 60.23* 6.25 8.06.91*

TABLE 22 S pyogenes Viability After Exposure to 12.5 μg/L of Penicillinwith Various Concentrations of DMSO Penicillin (μg/L) DMSO (%) S.pyogenes viability (%) 12.5 0 13.16 12.5 2.5 19.63 12.5 5.0 14.77* 12.58.0 6.43MSM in Combination with Penicillin

Similar experiments to those using DMSO were performed by combining MSMwith penicillin ranging from 3.125 to 12.5 μg/L. Results are shown inTables 23, 24, and 25. Synergy is indicated by an “*”. The reduction inbacterial viability due to MSM alone at 8% has been estimated to beapproximately 89%. As with DMSO, previously ineffective concentrationsof MSM and penicillin were effective in combination at reducingbacterial viability. When taken alone, no effect would be expected from3.13 μg/L penicillin with 2.5% MSM, however an 8% reduction in viabilityis detected (see Table 23). These effects are more pronounced with thecombination of 6.25 μg/L penicillin with MSM. For example, 5% MSM with6.25 μg/L penicillin would be expected to yield a 96% viable bacterialpopulation (see table 24). However, data indicate that viability wasreduced to about 17%, nearly an 80% reduction from expected results.Synergy was not detected when 12.5 μg/L penicillin was used, due to theefficacy of that concentration of penicillin alone. Thus, in someembodiments, MSM in concentrations between 2.5% and 8%, when combinedwith penicillin in concentrations of 3.125 to 6.25 μg/L result insynergistic decreases in bacterial viability. In several embodiments,previously ineffective concentrations of the compound are efficaciouswhen combined.

TABLE 23 S pyogenes Viability After Exposure to 3.13 μg/L of Penicillinwith Various Concentrations of MSM Penicillin (μg/L) MSM (%) S. pyogenesviability (%) 3.13 0 100 3.13 2.5 92.89* 3.13 5.0 78.31* 3.13 8.0 9.91*

TABLE 24 S pyogenes Viability After Exposure to 6.25 μg/L of Penicillinwith Various Concentrations of MSM Penicillin (μg/L) MSM (%) S. pyogenesviability (%) 6.25 0 100 6.25 2.5 90.11* 6.25 5.0 17.42* 6.25 8.0 10.77*

TABLE 25 S pyogenes Viability After Exposure to 12.5 μg/L of Penicillinwith Various Concentrations of MSM Penicillin (μg/L) MSM (%) S. pyogenesviability (%) 12.5 0 13.16 12.5 2.5 16.33 12.5 5.0 12.85 12.5 8.0 16.02

Example 8 Antibacterial Effects of DMSO and MSM in Combination withPenicillin on S. pyogenes

As with S. pneumonia, combinations of various concentrations of DMSO,MSM, and penicillin were evaluated for their effects on bacterialviability and possible synergistic activity as compared to MSM withpenicillin or DMSO with pencillin. Results are shown in Table 26.Synergy as compared to DMSO and penicillin is indicated by an “*” whilesynergy as compared to MSM and penicillin is indicated by an “ψ”. As canbe seen by the data in Table 26, substantial synergy was detected acrossthe various concentrations of compounds. Most combinations of DMSO andMSM exhibited a dose-response curve based on the concentration ofpenicillin used. Based on the efficiacy of 12.5 μg/L alone, it is notunexpected that combinations of this concentration of penicillin withDMSO and MSM should be more effective. Of interest, the previouslyineffective concentrations of pencillin are rendered effective in a dosedependent manner by combination with DMSO and MSM. For example, 2.5%DMSO with 5% MSM and 3.125 μg/L pencillin would be expected to reducebacterial viability to between 100% and 96% (when compared toDMSO+penicillin and MSM+penicillin, respectively). However, thecombination of all three reduced bacterial viability to about 19%. Theexpected results are similar for combinations with 6.25 μg/L penicillin,but the actual combination reduced bacterial viability even further, toabout 13%. Thus, in some embodiments, it appears that certainconcentrations of DMSO and MSM work as bacterial sensitizers, allowinglower concentrations of penicillin to function in a dose-dependentmanner, as if they were in fact higher concentrations of penicillin.Thus in some embodiments, certain combinations of DMSO, MSM, andpenicillin are advantageous because they render a previously inactiveset of individual compounds active against bacterial viability.

In some embodiments, simply increasing concentrations of the variouscompounds does not result larger reductions in bacterial viability. Forexample, the combination of 8% DMSO with 2.5% MSM and 3.125 μg/Lpenicillin appears to be more effective than 8% DMSO with 2.5% MSM and12.5 μg/L penicillin. Thus, in several embodiments, precise combinationsof each of the three compounds are used to maximize the reduction inbacterial viability. For example, in some embodiments, 2.5% to 5% DMSOin combination with 2.5, 5, or 20% MSM and with 3.125, 6.25, or 12.5μg/L penicillin results in synsergistic effects with respect tobacterial viability reduction. In one embodiment, 8% DMSO is used,wherein MSM is provided at 2.5% and penicillin is present at 3.125 μg/L.However, increasing either the MSM or penicillin when used with 8% DMSOreduces the overall efficacy of the combination.

TABLE 26 S. pneumonia Viability After Exposure to Various combinationsof DMSO, MSM, and Penicillin DMSO (%) MSM (%) Penicillin (μg/L) S.pyogenes viability (%) 2.5 2.5 3.125 91.74*,^(ψ) 2.5 2.5 6.2560.55*,^(ψ) 2.5 2.5 12.5 8.08*,^(ψ) 2.5 5 3.125 18.72*,^(ψ) 2.5 5 6.2513.38*,^(ψ) 2.5 5 12.5 9.41* 2.5 10 3.125 16.05*,^(ψ) 2.5 10 6.2511.78*,^(ψ) 2.5 10 12.5 11.77* 5 2.5 3.125 14.60*,^(ψ) 5 2.5 6.2510.44*,^(ψ) 5 2.5 12.5 9.55*,^(ψ) 8 2.5 3.125 9.55*,^(ψ) 8 2.5 6.2510.28^(ψ) 8 2.5 12.5 15.55^(ψ)

It will be understood by those of skill in the art that numerous andvarious modifications can be made without departing from the spirit ofthe present invention. Therefore, it should be clearly understood thatthe forms of the present invention are illustrative only and are notintended to limit the scope of the present invention.

What is claimed is:
 1. A method of sensitizing a drug resistantbacterial microorganism that does not cause tuberculosis to the drug towhich the microorganism is resistant, comprising: contacting themicroorganism with a formulation comprising 5-15% methylsulfonylmethane(MSM); and contacting the microorganism with the drug to which thebacterial microorganism is resistant, thereby sensitizing the drugresistant bacterial microorganism to the drug, wherein the microorganismis methicillin-resistant Staphylococcus aureus (MRSA) and the drug ismethicillin.
 2. The method of claim 1, wherein the microorganism iscontacted with the MSM before the microorganism is contacted with thedrug.
 3. The method of claim 1, wherein the microorganism is contactedwith the MSM at the same time that the microorganism is contacted withthe drug.
 4. The method of claim 1, wherein the microorganism iscontacted with a formulation comprising 7-15% MSM.
 5. The method ofclaim 1, wherein the formulation further comprises no more than 0.63%dimethylsulfoxide (DMSO).
 6. The method of claim 4, wherein theformulation comprises 7% MSM.
 7. The method of claim 4, wherein theformulation comprises 10% MSM.
 8. The method of claim 4, wherein theformulation comprises 15% MSM.
 9. The method of claim 1, whereincontacting the microorganism with the formulation comprising 5-15% MSM,the drug, or both, occurs is in a subject.
 10. The method of claim 9,wherein the microorganism is methicillin-resistant Staphylococcus aureus(MRSA).
 11. The method of claim 9, wherein the MSM is administeredtopically.
 12. The method of claim 9, wherein the MSM is administered byinhalation.